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Quantitative polarized light microscopy enables determination of optical retardation and azimuth of birefringent specimens and is a powerful tool for label-free imaging in the fields of biology and pathology. We have recently proposed a device for fast laser-scanning birefringence microscopy based on a near-infrared wavelength-swept laser and spectral encoding of polarization, resulting in a channeled spectrum generated during the wavelength-sweep of the laser and highly sensitive to optical retardation [Opt. Lett.49, 387 (2024)10.1364/OL.507576]. In this Letter, we propose its transposition to visible widefield imaging using a white light source and a high-order retarder for spectral encoding and a hyperspectral camera to record the channeled spectrum at each point of the image in parallel. The method proposed here allows for straightforward conversion of any widefield microscope into a highly sensitive and quantitative polarized light microscope.

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We have recently demonstrated a high-speed null polarimeter [Opt. Express30, 18889 (2022)10.1364/OE.454193OPEXFF1094-4087] based on passive polarization optics and using a fast swept-wavelength laser source. We report here its implementation in a laser-scanning microscope setup, enabling highly sensitive linear retardance imaging with a pixel dwell time of 10 µs. The instrument is also able to measure light depolarization induced by the sample. Images of biological samples, including cancerous tissue and cells, illustrate its performances.

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We recently developed a high speed null polarimeter [Opt. Express30, 18889 (2022)OPEXFF1094-408710.1364/OE.454193] based on passive polarization optics and a high speed wavelength swept laser source, enabling the measurement of linear retardance with 3.1µd e g/H z resolution within a minimum acquisition time of 10 µs, corresponding to a linear retardation of 8.6×10-9 λ/H z. The counterpart of high sensitivity lies in the systematic errors unlike Mueller polarimeters, which can be calibrated but which are much less sensitive. This paper focuses on the accuracy of this null polarimeter and provides hardware and numerical solutions to improve both linear retardance and azimuth measurements. Experiments and theoretical simulations are carried out to demonstrate the relevancy of these solutions.

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Null-polarimeters provide the best sensitivity to anisotropy measurements and so far have been developed for the detection of small optical activities. This paper revisits null polarimetry through an original configuration based on the concept of spectrally encoded light polarization, in order to measure, with unprecedented speed, either linear or circular retardance with the same degree of sensitivity . Using passive polarization optics and a high speed wavelength swept laser source, the achieved single-pass sensitivity was 55nrad/Hz and 45nrad/Hz for respectively linear and circular retardance considering a minimum acquisition time of 10 µs. Due to its compactness and rapidity, the method could be further implemented in laser scanning microscopes, which should be of great interest for revealing very low anisotropies in biological tissues.

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Gastric cancer (GC) is highly deadly. Three-dimensional (3D) cancer cell cultures, known as spheroids, better mimic tumor microenvironment (TME) than standard 2D cultures. Cancer-associated fibroblasts (CAF), a major cellular component of TME, promote or restrain cancer cell proliferation, invasion and resistance to drugs. We established spheroids from two human GC cell lines mixed with human primary CAF. Spheroid organization, analyzed by two-photon microscopy, showed CAF in AGS/CAF spheroids clustered in the center, but dispersed throughout in HGT-1/CAF spheroids. Such differences may reflect clonal specificities of GC cell lines and point to the fact that GC should be considered as a highly personalized disease.

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Among the multitude of optical polarization contrasts that can be observed in complex biological specimens, linear diattenuation (LD) imaging has received little attention. It is indeed challenging to image LD with basic polarizing microscopes because it is often relatively small in comparison with linear retardance (LR). In addition, interpretation of LD images is not straightforward when experiments are conducted in the visible range because LD can be produced by both dichroism and anisotropic scattering. Mueller polarimetry is a powerful implementation of polarization sensing able to differentiate and measure the anisotropies of specimens. In this article, near infrared transmission Mueller scanning microscopy is used to image LD in thin biological specimen sections made of various proteins with unprecedented resolution and sensitivity. The near infrared spectral range makes it possible to lower the contribution of dichroism to the total linear diattenuation in order to highlight anisotropic scattering. Pixel-by-pixel comparison of LD images with LR and multiphoton images demonstrates that LD is produced by under-resolved structures that are not revealed by other means, notably within the sarcomere of skeletal muscles. LD microscopy appears as a powerful tool to provide new insights into the macro-molecular organization of biological specimens at the sub-microscopic scale without labelling.

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In this paper, we report a generalized theoretical framework on spectrally encoded polarimeters to display in real time both linear and circular retardance as well as linear and circular diattenuation on the basis of a recent experimental work detailed in Sci. Rep.9, 3972 (2019)SRCEC32045-232210.1038/s41598-019-40467-z. The considered polarimeters use a broadband light source and wavelength-dependent retarders whose thicknesses are selected to guarantee minimal noise propagation and reduced data processing for real-time display of pure polarimetric effects.

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Mueller microscopes enable imaging of the optical anisotropic properties of biological or non-biological samples, in phase and amplitude, at sub-micrometre scale. However, the development of Mueller microscopes poses an instrumental challenge: the production of polarimetric parameters must be sufficiently quick to ensure fast imaging, so that the evolution of these parameters can be visualised in real-time, allowing the operator to adjust the microscope while constantly monitoring them. In this report, a full Mueller scanning microscope based on spectral encoding of polarization is presented. The spectrum, collected every 10 µs for each position of the optical beam on the specimen, incorporates all the information needed to produce the full Mueller matrix, which allows simultaneous display of all the polarimetric parameters, at the unequalled rate of 1.5 Hz (for an image of 256 × 256 pixels). The design of the optical blocks allows for the real-time display of linear birefringent images which serve as guidance for the operator. In addition, the instrument has the capability to easily switch its functionality from a Mueller to a Second Harmonic Generation (SHG) microscope, providing a pixel-to-pixel matching of the images produced by the two modalities. The device performance is illustrated by imaging various unstained biological specimens.

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Polarization-resolved second harmonic generation (P-SHG) microscopy is able to probe the sub-micrometer structural organization of myosin filaments within skeletal muscle. In this study, P-SHG microscopy was used to analyze the structural consequences of sepsis, which is the main cause of the critical illness polyneuromyopathy (CIPNM). Experiments conducted on two populations of rats demonstrated a significant difference of the anisotropy parameter between healthy and septic groups, indicating that P-SHG microscopy is promising for the diagnosis of CIPNM. The difference, which can be attributed to a change of myosin conformation at the sub-sarcomere scale, cannot be evidenced by classical SHG imaging.

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A new high-speed second-harmonic generation (SHG) polarimetric method is reported. It is based on the spectral analysis of the SHG radiation emitted by a nonlinear medium excited with circularly polarized femtosecond laser pulses. The setup uses only passive components for polarization encoding and a fast spectrometer for spectral analysis. The method is validated on a z-cut quartz plate, then on a collagen-rich biological tissue with a view to single image polarization-resolved SHG microscopy.

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A full Mueller polarimeter was implemented on a commercial laser-scanning microscope. The new polarimetric microscope is based on high-speed polarization modulation by spectral coding using a wavelength-swept laser as a source. Calibration as well as estimation of the measurement errors of the device are reported. The acquisition of Mueller images at the speed of a scanning microscope is demonstrated for the first time. Mueller images of mineral and biological samples illustrate this new polarimetric microscopy.

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BACKGROUND AND AIMS: Statins are prescribed for their preventative effects within atherosclerosis development. To our knowledge, no study focusing on very low-dose (non-hypolipidemic effect) and long-term atorvastatin treatment in vivo was available. Our aim was to assess the effect of such atorvastatin treatment on the mechanical and functional characteristics of arteries in the context of primary prevention. METHODS: An atorvastatin treatment (2.5 mg/kg/day) was tested against controls on 34 male 3 to 12 month-old WHHL rabbits. No effect on total cholesterol, triglycerides, HDL or LDL was observed. The arterial stiffness was evaluated on vigil animals by pulse wave velocity (PWV) measurement. Then, in vitro measurements were made to evaluate (1) the endothelial and vascular smooth muscle function, (2) the elasticity of the arterial wall and (3) the composition in collagen and elastin in the aorta. RESULTS: The PWV increasing observed with age in control group was canceled by treatment, creating a significance difference between groups at 12 months (5.17 ± 0.50 vs 2.14 ± 0.34 m s(-1) in control and treated groups respectively). Vasoreactivity modifications can't explain this result but maintain of elasticity with treatment in large arteries was confirm by a static tensile test. A first possible explanation is the change of wall composition with treatment, validated by the percentage of elastin at 12 months, 4.4% lower in the control group compared to the treated group (p < 0.05). CONCLUSIONS: This study shows that a non-hypocholesterolemic statin treatment could improve vessel elasticity in the atherosclerotic WHHL model. The great novelty of this work is the vessel wall composition changing associated. This first approach in animal opens the reflection on the use of these low doses in humans. This could be interesting in the context of arterial stiffening with aging, non-hyperlipidemic atherosclerosis or with cholesterol reduce by another therapy or lifestyle modification.

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In this paper we analyze a fibrosis scoring method based on measurement of the fibrillar collagen area from second harmonic generation (SHG) microscopy images of unstained histological slices from human liver biopsies. The study is conducted on a cohort of one hundred chronic hepatitis C patients with intermediate to strong Metavir and Ishak stages of liver fibrosis. We highlight a key parameter of our scoring method to discriminate between high and low fibrosis stages. Moreover, according to the intensity histograms of the SHG images and simple mathematical arguments, we show that our area-based method is equivalent to an intensity-based method, despite saturation of the images. Finally we propose an improvement of our scoring method using very simple image processing tools.

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A new setup is proposed to perform high-speed Mueller polarimetry by spectral coding of polarization in a reflection configuration. The system uses a swept laser source and a photodiode, which results in a simple optical setup that allows measurement of Mueller matrices at 100 kHz repetition rate. A special focus is made on the influence of the cube beam splitter polarimetric response, which is essential to measurements in a reflection configuration. The instrument is first validated on reference samples for single-point measurements, and the effect of a proper system calibration is also demonstrated on polarimetric images. The device is intended to be implemented within a laser scanning microscope to perform multimodal imaging (confocal/multiphoton and Mueller polarimetry).

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In this paper, we propose a new and simple method based on two-photon excitation fluorescence (TPEF) microscopy to measure the scattering coefficient µ(s) of thick turbid media. We show, from Monte Carlo simulations, that µ(s) can be derived from the axial profile of the ratio of the TPEF signals epi-collected by the confocal and the non-descanned ports of a scanning microscope, independently of the anisotropy factor g and of the absorption coefficient µ(a) of the medium. The method is validated experimentally on tissue-mimicking optical phantoms, and is shown to have potential for imaging the scattering coefficient of heterogeneous media.

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Experimental results are presented related to the dynamic behaviour of Polymer Stabilized Ferro-electric Liquid Crystal (PSFLC) samples under external applied electric field, using Snap-shot Mueller Matrix Polarimetry (SMMP) and Mueller Matrix (MM) formalism. Different polarimetric coefficients are simultaneously extracted from each channeled spectrum measured with this full-optical SMMP technique. The impact of the concentration of polymer present into the liquid crystal cell on this dynamic behaviour is studied, permitting a direct and quick characterisation of the material. The results obtained for PSFLC are compared with those already measured for pure Surface Stabilized Ferro-electric Liquid Crystal (SSFLC) samples, which correspond to a 0% concentration in polymer.

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An experimental Mueller matrix polarimeter is used to quantify human liver fibrosis by measuring retardance and depolarization of thin biopsies. The former parameter is sensitive to fibrillar collagen, the latter is specifically sensitive to fibrillar collagen around blood vessels, which is not significant for liver fibrosis diagnosis. By using depolarization like a filter, retardance distribution enables distinguishing between disease stages and limits the high degree of observer discrepancy.

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An experimental snapshot Mueller matrix polarimeter based on wavelength polarization coding is used to get a time-resolved description of electric-field-induced fast transition within a ferroelectric liquid-crystal cell. The parameters extracted from experimental Mueller matrices are linked to the molecule director distribution to further determine the average trajectory and the collective behavior of these molecules while they switch over to another state.

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We describe a new setup for a snapshot Mueller matrix polarimeter (SMMP). It relies on the separation and orthogonal polarization of two light beams by a Wollaston prism located at the setup output. The simultaneous treatment of the two spectra allows an enhancement of accuracy for real-time measurements through reduction of the effects caused by random noise and systematic errors. Moreover, it gives insight into the nonuniform spectral response of the medium under study. Experimental results support the feasibility of the proposed technique.

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Systematic errors specific to a snapshot Mueller matrix polarimeter are studied. Their origins and effects are highlighted, and solutions for correction and stabilization are proposed. The different effects induced by them are evidenced by experimental results acquired with a given setup and theoretical simulations carried out for more general cases. We distinguish the errors linked to some imperfection of elements in the experimental setup from those linked to the sample under study.